Attempts have been made to fabricate nanofibrous scaffolds to mimic the chemical composition and structural properties of extracellular matrix (ECM) for tissue/organ regeneration. Nanofibers with various patterns have been successfully produced from synthetic and natural polymers through a relatively simple technique of electrospinning. The resulting patterns can mimic some of the diverse tissue-specific orientation and three-dimensional (3D) fibrous structure. Studies on cell-nanofiber interactions have revealed the importance of nanotopography on cell adhesion, proliferation and differentiation. Our recent data showed that hematopoietic stem cells (HSCs) as well as mesenchymal stem cells (MSCs) can rapidly and effectively attached to the functionalized nanofibers. Mineralized 3D nanofibrous scaffold with bone marrow derived MSCs has been applied for bone tissue engineering. The use of injectable nanofibers for cardiac tissue engineering applications is attractive as they allow for the encapsulation of cardiomyocytes/MSCs as well as bioactive molecules for the repair of myocardial infarction. Duplicate 3D heart helix microstructure by the nanofibrous cardiac patch might provide functional support for infarcted myocardium. Furthermore, clinical applications of electrospun nanofibers for regenerative medicine are highly feasible due to the ease and flexibility of fabrication with the cost-effective method of making nanofibers.